裸鼠肝部分切除术后肝癌血管内皮生长因子及微血管密度变化的研究

目的研究肝部分切除后裸鼠肝癌组织微血管密度(MVD)及血管内皮生长因子(VEGF)的表达,以探讨其促进肿瘤生长的机制.方法采用切除裸鼠肝左叶和中叶观察肝肿瘤生长情况,以免疫组化和图像分析检测肝癌组织MVD、VEGF的表达.结果肝部分切除后,肿瘤组织MVD和VEGF表达明显增强,肿瘤组织重量和体积明显增加.结论肝部分切除后肿瘤组织VEGF表达增强,肿瘤新生血管形成增多可能为其促进肿瘤生长的机制.     

体外化疗药敏试验系统联合流式细胞仪在胃肠肿瘤化疗中的应用

目的：探讨体外化疗药敏试验系统（ATP－TCA系统）联合流式细胞仪（FCM）在胃肠肿瘤化疗中的应用。方法：取36例胃癌、结直肠癌的手术标本行ATP－TCA试验和FCM P170检测，并分析两结果之间的关系。结果：ATP－TCA的可评估率为88．1％，10种化疗药物的敏感率分别为：氟尿嘧啶15．6％，顺铂18．8％，足叶乙甙21．9％，草酸铂25．0％，丝裂霉素31．3％，表阿霉素37．5％，诺消灵40．6％，开普拓43．8％，健择53．1％，泰素56．3％，P170检测阳性率为43．8％，丝裂霉素，表阿霉素，诺消灵的敏感率在P170阴性表达时高，开普拓，健择，泰素在P170阳性和阴性表达时的敏感率均高，且差异无显性意义（P＞0．05），结论：ATP－TCA法联合FCM可较好地用于胃肠肿瘤的化疗药物的筛选，开普拓，健择，泰素可用于P170阳性表达的患。     

术前肠内营养作为肠道准备方式对老年大肠癌术后早期康复的观察

目的探讨采用肠内营养作为肠道准备对老年大肠癌术后早期康复的影响。方法回顾性分析56例老年结直肠癌患者的临床资料,按照术前肠道准备的不同分为肠内营养组和对照组。肠内营养组术前口服肠内营养混悬液,对照组则采用传统治疗方法(普通流质食物),观察比较两组疗效及术后并发症。结果两组手术时间、总住院时间等相关指标接近,均未发现吻合口漏及腹腔感染等严重并发症。肠内营养组患者术中肠道清洁度及术后营养指标均优于对照组,差异有统计学意义(P0.05),两组患者肠道准备期间不良反应无显著差异。结论老年结直肠癌患者肠道术前准备使用肠内营养安全可行,避免传统方法的缺点,且为手术期效果良好。     

受翻译调节的肿瘤蛋白在PRL-3促进结肠癌转移中的作用

目的: 研究受翻译调节的肿瘤蛋白(TCTP)在肝再生磷酸酶-3(PRL-3)促进结肠癌细胞增殖、迁移和侵袭中的作用。方法: 构建载体pAcGFP-C3-PRL-3及pAcGFP-C3并分别转染至结肠癌LoVo细胞中,获得稳定表达PRL-3的LoVo-PRL-3细胞及对照细胞LoVo-control。Western blotting及real-time PCR 检测2种细胞PRL-3及TCTP表达。设计并合成特异性干扰TCTP mRNA的siRNA序列(TCTP-siRNA)和阴性对照序列(control-siRNA),并将siRNA瞬时转染至LoVo-PRL-3细胞。 在转染siRNA后24 h、48 h和72 h,Western blotting及real-time PCR 检测LoVo-PRL-3细胞TCTP表达。 CCK8-8和Transwell方法检测LoVo-control、LoVo-PRL-3及转染TCTP-siRNA和转染control-siRNA的LoVo-PRL-3细胞间增殖、迁移和侵袭能力差异。结果: 转染PRL-3可以显著上调LoVo细胞TCTP mRNA和蛋白的表达(P＜0.05)。TCTP-siRNA在转染后的24 h、48 h和72 h可以有效抑制LoVo-PRL-3细胞TCTP mRNA表达(P＜0.01),同样TCTP蛋白在转染后的48 h和72 h也显著受到抑制(P＜0.01)。转染PRL-3能显著促进LoVo细胞增殖、迁移和侵袭能力(P＜0.05),然而siRNA干扰抑制上调的TCTP表达后又能显著抑制LoVo-PRL-3细胞增殖、迁移和侵袭能力(P＜0.05)。结论: PRL-3通过上调TCTP表达促进结肠癌细胞增殖、迁移和侵袭。用siRNA靶向抑制TCTP表达可能是预防和治疗结肠癌转移的有效手段。     

不同细胞因子诱导对树突状细胞体外分化的影响

目的 观察不同细胞因子诱导对于体外培养的树突状细胞(DC)免疫刺激活性的影响.方法 通过1000 U/ml人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)和500 U/ml白细胞介素(IL)-4体外诱导的人单个核细胞来源的DC,经1000 U/ml肿瘤坏死因子(TNF)-α、1 mg/L脂多糖(LPS)、1 mmol/L丁酸钠、细胞因子鸡尾酒法诱导成熟,分别以流式细胞仪、FITC-Dxtran内吞检测、混合淋巴细胞反应(MLR)、酶联免疫吸附试验(ELISA)检测DC的表面标志、内吞能力、刺激淋巴细胞增殖能力和IL-12分泌的变化.结果 鸡尾酒法诱导下的DC成熟标志显著上调,内吞能力减弱186.74±38.66,刺激淋巴细胞增殖能力增强18.23 ±2.22,并且IL-12的分泌能力增加(656.18±38.52)ng/L,说明其显著促进DC成熟.而丁酸钠诱导下的各项成熟指标均下调,导致DC免疫刺激源性减弱.结论 细胞因子鸡尾酒法是诱导DC成熟的最佳方法;而丁酸钠可以抑制DC成熟,改变DC的免疫状态.

Abstract：

Objective To investigate the immune effect of different cytokines on immature dendritic cells (DC) in vitro. Methods The human monocyte-derived DCs were induced in the presence of recombinant human GM-CSF (rhGM-CSF, 1000 U/ml) and interleukin (IL)-4 (500 U/ml). The effects of tumor necrosis factor (TNF) -α (1000 U/ml) , lipopolysaccharide (LPS) (1 mg/L), the cocktail of cytokines (TNF-α, IL-6, IL-1β, PGE2) , and sodium butyrate (1 mmol/L) on the DCs were detected by flow cytometry (FCM), endocytic activity, T cells stimulatory proliferation capacity, and IL-12 production. Results The cocktail of cytokines could up-regulate the major histocompatibility complex (MHC) class Ⅱ and costimulatory molecules of DCs, decrease the endocytic activity 186. 74 ±38. 66, induce a stage of Tcell stimulation 18. 23 ± 2. 22 and promote the T helper cell type 1-skewing factor IL-12 production (656. 18 ±38. 52) ng/L. But the sodium butyrate induced reverse result on DCs compared to cocktail of cytokines. Conclusion The most effective method for inducing dendritic cells maturation was cocktail of cytokines. The sodium butyrate could inhibit the development of mature DCs, resulting in impaired DC function, and modify the outcome of the subsequent immune response.     

猪远端胃大部分切除术后合并门静脉高压症模型中门脉血流系统的变化

目的 建立猪远端胃大部分切除术后合并门静脉高压症模型,观察该模型门脉系统的血流动力学变化.方法 将26只家猪随机分为3组,实验组(EG,10只)一期行远端胃大部分切除、二期行门静脉部分缩窄术;门静脉高压组(PG,10只)一期行开关腹、二期行门静脉部分缩窄术;假手术组(SG,6只)一、二期均行开关腹.3组均测定门静脉压力,二期手术后两月行CTA及腹腔干、门静脉系统血管铸型.结果 与假手术组比较,实验组、门静脉高压组术后均形成了门静脉高压.实验组与门静脉高压组术后及术后两个月门静脉压力差异无统计学意义(P＞0.05).远端胃大部分切除术后合并门静脉高压症,胃的血供主要由胃左动静脉及胃网膜左动静脉提供,没有形成新的侧支循环.结论 远端胃大部分切除术后合并门静脉高压症,不宜行贲门周围血管离断术,治疗上需对患者的门腔系统行血管成像检查,根据情况选择适当的治疗方式.     

抗原负载的树突状细胞体外诱导抗肿瘤免疫的研究

目的 观察抗原负载的树突状细胞(DCs)体外诱导抗肿瘤免疫效应,探讨树突状细胞肿瘤疫苗的制备方法 .方法 以细胞因子粒细胞-巨噬细胞集落刺激因子(GM-CSF)和白细胞介素(IL)-4体外诱导人单核细胞来源的树突状细胞,第6天加入肝癌细胞BeL-7402的冻融抗原并以联合细胞因子鸡尾酒法[肿瘤坏死因子(TNF-α)+IL-6+IL-1β+前列腺素E2(PGE2)]诱导成熟,对照组仅以鸡尾酒法诱导成熟.24 h后收获DCs以流式细胞仪检测其成熟表型CD80、CD83、CD86和LHA-DR,酶联免疫吸附试验(ELISA)法检测其IL-12的分泌,噻唑蓝(MTT)比色法检测其刺激淋巴细胞增殖活性,乳酸脱氢酶释放实验检测其诱导的免疫效应细胞对肝癌细胞的特异性细胞毒作用.结果 联合细胞因子可诱导的DCs成熟和IL-12的分泌(P＜0.05),成熟的DCs有较强的刺激淋巴细胞增殖能力;抗原负载组DCs可诱导效应细胞对肝癌细胞BeL-7402的特异性杀伤作用(P＜0.05).结论 抗原负载的DCs可体外诱导特异性抗肿瘤免疫效应,提示以抗原负载的树突状细胞作为肿瘤免疫治疗的方法 是可行的.     

裸鼠肝部分切除后肝癌组织基质金属蛋白酶-2及其抑制剂表达

［目的］观察肝部分切除后裸鼠肝癌组织基质金属蛋白酶-２(MMP-２)及其抑制剂(TIMP-２)表达的变化，以探讨肝部分切除促肿瘤生长的机制。［方法］２０只裸鼠随机分成２组：对照组和肝切除组，每组１０只，对照组仅将人肝癌组织种植于右肝叶，肝切除组切除肝左叶和中叶后于右肝叶种植人肝癌组织，２组动物于３５天后处死，检测肿瘤大小和重量，采用免疫组化和图像分析法检测肝癌组织微血管密度(MVD)、MMP-２和TIMP-２的表达。［结果］肝切除组肿瘤重量和体积、MVD和MMP-２表达较对照组明显增加(P＜０．０５)，TIMP-２则明显减低(P＜０．０５)。［结论］肝部分切除后MMP-２表达上调、TIMP-２表达下调导致肿瘤新生血管形成增多，可能为其促肿瘤生长的机制。     

结直肠癌患者循环肿瘤细胞监测的研究进展

尽管近年来结直肠癌手术、化疗、放疗的疗效较前已有明显改善,但仍有15%~20%的结直肠癌患者在诊断时就已发现肝转移,而其余60%亦在后续的治疗过程中发现复发或转移。而外周血中循环肿瘤细胞(CTCs)的检测作为近年来临床研究的热点,在评估治疗癌症的疗效及判断预后等方面有着巨大的潜力。本文就CTCs的分离富集、检测技术及结直肠癌患者中CTCs检测的临床意义进行综述。     

Proteomic analysis identifies translationally controlled tumor protein as a mediator of phosphatase of regenerating liver-3-promoted proliferation, migration and invasion in human colon cancer cells

Background  Considerable evidence suggests that phosphatase of regenerating liver-3 (PRL-3) plays multiple roles in cancer metastasis; however, the molecular mechanisms remain largely unknown. The aim of this study was to identify proteins associated with PRL-3-promoted colon cancer metastasis, by comparative proteomic analysis.

Methods  Proteomes of human colon cancer LoVo cells transfected with PRL-3 gene (LoVo-PRL-3) or empty vector PAcGFP-C3 (LoVo-control) were compared using 2D gel electrophoresis. Proteins that varied significantly in concentration were selected and identified using mass spectrometry. Expression of translationally controlled tumor protein (TCTP) mRNA and protein in LoVo-PRL-3 and LoVo-control cells was detected by real-time PCR and Western blotting. Small interfering RNA (siRNA) targeting TCTP was used for silencing TCTP expression in LoVo-PRL-3 cells. Functional significance of TCTP in PRL-3-promoted colon cancer cell proliferation, migration and invasion was investigated by Cell Counting Kit-8 assay and transwell chamber.

Results  Seventeen proteins displaying significant and reproducible differences between LoVo-PRL-3 and LoVo-control cells were identified. Ten proteins were upregulated and seven were downregulated in LoVo-PRL-3 cells when compared with LoVo-control cells. Eight identified proteins are associated with distinct steps of tumor metastasis: ubiquitin-like protein ISG15, interleukin-18, TCTP, serpin B5, annexin A3, macrophage-capping protein, ATP-dependent RNA helicase DDX3X, and cathepsin D. Real-time PCR and Western blotting results showed that both TCTP mRNA and protein were significantly increased in LoVo-PRL-3 cells compared to LoVo-control cells. Transfection with TCTP siRNA significantly reduced the expression of both mRNA and protein levels of TCTP in LoVo-PRL-3 cells. Knockdown of TCTP by siRNA inhibited PRL-3-promoted proliferation, migration and invasion of LoVo-PRL-3 cells.

Conclusion  Our results imply that TCTP might be a mediator of PRL-3-promoted proliferation, migration and invasion of human colon cancer cells.